BMRB

Biological Magnetic Resonance Data Bank


A Repository for Data from NMR Spectroscopy on Proteins, Peptides, Nucleic Acids, and other Biomolecules
Member of WWPDB

BMRB Entry 16659

Title: Assignment and structural characterization of intrinsically disordered CDK inhibitor phosphoSic1 from yeast   PubMed: 20399186

Authors: Mittag, Tanja; Choy, Wing-Yiu; Marsh, Joseph; Orlicky, Stephen; Grishaev, Alexander; Lin, Hong; Sicheri, Frank; Tyers, Mike; Forman-Kay, Julie

Citation: Mittag, Tanja; Marsh, Joseph; Grishaev, Alexander; Orlicky, Stephen; Lin, Hong; Sicheri, Frank; Tyers, Mike; Forman-Kay, Julie. "Structure/function implications in a dynamic complex of the intrinsically disordered Sic1 with the Cdc4 subunit of an SCF ubiquitin ligase."  Structure (Cambridge, MA, U. S.) 18, 494-506 (2010).

Assembly members:
pSic1, polymer, 92 residues, 10195 Da.
pSic1_G-1X, polymer, 92 residues, Formula weight is not available
pSic1_N21X, polymer, 92 residues, Formula weight is not available
pSic1_S38X, polymer, 92 residues, Formula weight is not available
pSic1_N64X, polymer, 92 residues, Formula weight is not available
pSic1_P83X, polymer, 92 residues, Formula weight is not available
pSic1_T90X, polymer, 92 residues, Formula weight is not available

Natural source:   Common Name: baker   Taxonomy ID: 4932   Superkingdom: not available   Kingdom: not available   Genus/species: Eukaryota Fungi

Experimental source:   Production method: recombinant technology   Host organism: Escherichia coli

Entity Sequences (FASTA):
pSic1: GSMTPSXPPRSRGTRYLAQP SGNTSSSALMQGQKXPQKPS QNLVPVXPSTTKSFKNAPLL APPNSNMGMTXPFNGLTXPQ RXPFPKSSVKRT
pSic1_G-1X: XSMTPSXPPRSRGTRYLAQP SGNTSSSALMQGQKXPQKPS QNLVPVXPSTTKSFKNAPLL APPNSNMGMTXPFNGLTXPQ RXPFPKSSVKRT
pSic1_N21X: GSMTPSXPPRSRGTRYLAQP SGXTSSSALMQGQKXPQKPS QNLVPVXPSTTKSFKNAPLL APPNSNMGMTXPFNGLTXPQ RXPFPKSSVKRT
pSic1_S38X: GSMTPSXPPRSRGTRYLAQP SGNTSSSALMQGQKXPQKPX QNLVPVXPSTTKSFKNAPLL APPNSNMGMTXPFNGLTXPQ RXPFPKSSVKRT
pSic1_N64X: GSMTPSXPPRSRGTRYLAQP SGNTSSSALMQGQKXPQKPS QNLVPVXPSTTKSFKNAPLL APPNSXMGMTXPFNGLTXPQ RXPFPKSSVKRT
pSic1_P83X: GSMTPSXPPRSRGTRYLAQP SGNTSSSALMQGQKXPQKPS QNLVPVXPSTTKSFKNAPLL APPNSNMGMTXPFNGLTXPQ RXPFXKSSVKRT
pSic1_T90X: GSMTPSXPPRSRGTRYLAQP SGNTSSSALMQGQKXPQKPS QNLVPVXPSTTKSFKNAPLL APPNSNMGMTXPFNGLTXPQ RXPFPKSSVKRX

Data sets:
Data typeCount
13C chemical shifts171
15N chemical shifts75
1H chemical shifts75
heteronuclear NOE values74
residual dipolar couplings63
T1 relaxation values74
T2 relaxation values74

Additional metadata:

  • Assembly
  • Samples and Experiments
  • Software
  • Spectrometers
  • Hide all

Assembly:

Entity Assembly IDEntity NameEntity ID
1pSic11

Entities:

Entity 1, pSic1 92 residues - 10195 Da.

The first two residues represent a non-native artifact from cleavage of an affinity tag and are therefore numbered -1 and 0. Residue M1 is the actual beginning of the native Sic1 sequence. The construct used represents the first 90 residues of Sic1, the so-called targeting region. Residues 5, 33 and 45 are phosphoThr, 69,76 and 80 phosphoSer.

1   GLYSERMETTHRPROSERTPOPROPROARG
2   SERARGGLYTHRARGTYRLEUALAGLNPRO
3   SERGLYASNTHRSERSERSERALALEUMET
4   GLNGLYGLNLYSTPOPROGLNLYSPROSER
5   GLNASNLEUVALPROVALTPOPROSERTHR
6   THRLYSSERPHELYSASNALAPROLEULEU
7   ALAPROPROASNSERASNMETGLYMETTHR
8   SEPPROPHEASNGLYLEUTHRSEPPROGLN
9   ARGSEPPROPHEPROLYSSERSERVALLYS
10   ARGTHR

Samples:

sample_1: pSic1, [U-13C; U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_2: pSic1, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_3: pSic1, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; PEG(C12E5)/hexanol 0.08 v/v; H2O 90%; D2O 10%

sample_4: pSic1_G-1X, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_5: pSic1_N21X, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_6: pSic1_S38X, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_7: pSic1_N64X, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_8: pSic1_P83X, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_9: pSic1_T90X, [U-15N], 0.5 mM; potassium phosphate 50 mM; sodium chloride 150 mM; sodium azide 0.02%; EDTA 1 mM; DSS 1 mM; H2O 90%; D2O 10%

sample_conditions_1: ionic strength: 0.2 M; pH: 7.0; pressure: 1 atm; temperature: 278 K

Experiments:

NameSampleSample stateSample conditions
2D 1H-15N HSQCsample_2isotropicsample_conditions_1
3D CBCA(CO)NHsample_1isotropicsample_conditions_1
3D HNCACBsample_1isotropicsample_conditions_1
3D C(CO)NHsample_1isotropicsample_conditions_1
3D HNCOsample_1isotropicsample_conditions_1
pseudo3D 15N T1sample_2isotropicsample_conditions_1
pseudo3D 15N T1rhosample_2isotropicsample_conditions_1
pseudo3D 1H-15N hetNOEsample_2isotropicsample_conditions_1
2D 1H-15N IPAPsample_3anisotropicsample_conditions_1
pseudo3D NH R2 for PREsample_4isotropicsample_conditions_1
pseudo3D NH R2 for PREsample_5isotropicsample_conditions_1
pseudo3D NH R2 for PREsample_6isotropicsample_conditions_1
pseudo3D NH R2 for PREsample_7isotropicsample_conditions_1
pseudo3D NH R2 for PREsample_8isotropicsample_conditions_1
pseudo3D NH R2 for PREsample_9isotropicsample_conditions_1

Software:

ENSEMBLE, Marsh, Choy, Forman-Kay - ensemble generation

NMRPipe, Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax - processing

NMRView, Johnson, One Moon Scientific - chemical shift assignment

FuDA, Kristensen and Hansen - peak integration, RDC analysis

NMR spectrometers:

  • Varian INOVA 500 MHz
  • Varian INOVA 800 MHz

Related Database Links:

UNP P38634